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Journal of Biomedical Optics Jun 2024Three-dimensional quantitative phase imaging (QPI) has rapidly emerged as a complementary tool to fluorescence imaging, as it provides an objective measure of cell...
SIGNIFICANCE
Three-dimensional quantitative phase imaging (QPI) has rapidly emerged as a complementary tool to fluorescence imaging, as it provides an objective measure of cell morphology and dynamics, free of variability due to contrast agents. It has opened up new directions of investigation by providing systematic and correlative analysis of various cellular parameters without limitations of photobleaching and phototoxicity. While current QPI systems allow the rapid acquisition of tomographic images, the pipeline to analyze these raw three-dimensional (3D) tomograms is not well-developed. We focus on a critical, yet often underappreciated, step of the analysis pipeline that of 3D cell segmentation from the acquired tomograms.
AIM
We report the CellSNAP (Cell Segmentation via Novel Algorithm for Phase Imaging) algorithm for the 3D segmentation of QPI images.
APPROACH
The cell segmentation algorithm mimics the gemstone extraction process, initiating with a coarse 3D extrusion from a two-dimensional (2D) segmented mask to outline the cell structure. A 2D image is generated, and a segmentation algorithm identifies the boundary in the plane. Leveraging cell continuity in consecutive -stacks, a refined 3D segmentation, akin to fine chiseling in gemstone carving, completes the process.
RESULTS
The CellSNAP algorithm outstrips the current gold standard in terms of speed, robustness, and implementation, achieving cell segmentation under 2 s per cell on a single-core processor. The implementation of CellSNAP can easily be parallelized on a multi-core system for further speed improvements. For the cases where segmentation is possible with the existing standard method, our algorithm displays an average difference of 5% for dry mass and 8% for volume measurements. We also show that CellSNAP can handle challenging image datasets where cells are clumped and marred by interferogram drifts, which pose major difficulties for all QPI-focused AI-based segmentation tools.
CONCLUSION
Our proposed method is less memory intensive and significantly faster than existing methods. The method can be easily implemented on a student laptop. Since the approach is rule-based, there is no need to collect a lot of imaging data and manually annotate them to perform machine learning based training of the model. We envision our work will lead to broader adoption of QPI imaging for high-throughput analysis, which has, in part, been stymied by a lack of suitable image segmentation tools.
Topics: Humans; Image Processing, Computer-Assisted; Imaging, Three-Dimensional; Quantitative Phase Imaging; Algorithms; Optical Imaging
PubMed: 38638450
DOI: 10.1117/1.JBO.29.S2.S22706 -
Journal of Biomedical Optics Apr 2024Light-sheet fluorescence microscopy (LSFM) has emerged as a powerful and versatile imaging technique renowned for its remarkable features, including high-speed 3D...
SIGNIFICANCE
Light-sheet fluorescence microscopy (LSFM) has emerged as a powerful and versatile imaging technique renowned for its remarkable features, including high-speed 3D tomography, minimal photobleaching, and low phototoxicity. The interference light-sheet fluorescence microscope, with its larger field of view (FOV) and more uniform axial resolution, possesses significant potential for a wide range of applications in biology and medicine.
AIM
The aim of this study is to investigate the interference behavior among multiple light sheets (LSs) in LSFM and optimize the FOV and resolution of the light-sheet fluorescence microscope.
APPROACH
We conducted a detailed investigation of the interference effects among LSs through theoretical derivation and numerical simulations, aiming to find optimal parameters. Subsequently, we constructed a customized system of multi-LSFM that incorporates both interference light sheets (ILS) and noninterference light-sheet configurations. We performed beam imaging and microsphere imaging tests to evaluate the FOV and axial resolution of these systems.
RESULTS
Using our custom-designed light-sheet fluorescence microscope, we captured the intensity distribution profiles of both interference and noninterference light sheets (NILS). Additionally, we conducted imaging tests on microspheres to assess their imaging outcomes. The ILS not only exhibits a larger FOV compared to the NILS but also demonstrates a more uniform axial resolution.
CONCLUSIONS
By effectively modulating the interference among multiple LSs, it is possible to optimize the intensity distribution of the LSs, expand the FOV, and achieve a more uniform axial resolution.
Topics: Microscopy, Fluorescence; Microspheres; Photobleaching
PubMed: 38629030
DOI: 10.1117/1.JBO.29.4.046501 -
Bioorganic & Medicinal Chemistry May 20245-Aminolevulinic acid (ALA) and its derivatives, serving as the endogenous precursor of the photosensitizer (PS) protoporphyrin IX (PpIX), successfully applied in tumor...
5-Aminolevulinic acid (ALA) and its derivatives, serving as the endogenous precursor of the photosensitizer (PS) protoporphyrin IX (PpIX), successfully applied in tumor imaging and photodynamic therapy (PDT). ALA and its derivatives have been used to treat actinic keratosis (AK), basal cell carcinoma (BCC), and improve the detection of superficial bladder cancer. However, the high hydrophilicity of ALA and the conversion of PpIX to heme have limited the accumulation of PpIX, hindering the efficiency and potential application of ALA-PDT. This study aims to evaluate the PDT activity of three rationally designed series of ALA-HPO prodrugs, which were based on enhancing the lipophilicity of the prodrugs and reducing the labile iron pool (LIP) through HPO iron chelators to promote PpIX accumulation. Twenty-four ALA-HPO conjugates, incorporating amide, amino acid, and ester linkages, were synthesized. Most of the conjugates, exhibited no dark-toxicity to cells, according to bioactivity evaluation. Ester conjugates 19a-g showed promoted phototoxicity when tested on tumor cell lines, and this increased phototoxicity was strongly correlated with elevated PpIX levels. Among them, conjugate 19c emerged as the most promising (HeLa, IC = 24.25 ± 1.43 μM; MCF-7, IC = 43.30 ± 1.76 μM; A375, IC = 28.03 ± 1.00 μM), displaying superior photodynamic anticancer activity to ALA (IC > 100 μM). At a concentration of 80 μM, the fluorescence intensity of PpIX induced by compound 19c in HeLa, MCF-7, and A375 cells was 18.9, 5.3, and 2.8 times higher, respectively, than that induced by ALA. In conclusion, cellular phototoxicity showed a strong correlation with intracellular PpIX fluorescence levels, indicating the potential application of ALA-HPO conjugates in ALA-PDT.
Topics: Humans; Aminolevulinic Acid; Photosensitizing Agents; Photochemotherapy; Antineoplastic Agents; Structure-Activity Relationship; Drug Screening Assays, Antitumor; Molecular Structure; Pyridones; Cell Line, Tumor; Protoporphyrins; Cell Proliferation; Dose-Response Relationship, Drug; Cell Survival; Prodrugs
PubMed: 38626642
DOI: 10.1016/j.bmc.2024.117726 -
Georgian Medical News Feb 2024The Nitrosogenesis of skin cancer is a modern newly introduced concept in medicine, mainly concerning melanoma, but also keratinocytic cancers such as basal cell...
PERIOCULAR HIGH RISK BCCS AFTER ADDITIONAL/PARALLEL INTAKE OF TORASEMIDE, MOXONIDINE AND MIRABEGRON: IMPORTANT LINKS TO SKIN CANCER RELATED (PHOTO-) NITROSOGENESIS IN THE CONTEXT OF PHARMACO-ONCOGENESIS.
The Nitrosogenesis of skin cancer is a modern newly introduced concept in medicine, mainly concerning melanoma, but also keratinocytic cancers such as basal cell carcinoma. The nitroso-contamination of more than 300 drugs worldwide and the permanent (relatively short-term) intake of mutagen-contaminated drugs could create serious prerequisites for the development of skin cancer. Retrospective but also prospective analyses following potentially contaminated polymedication with a heterogeneous type of nitrosamines in real patients are indicative of a causal connection rather than a sporadic association between 1) intake of a possibly nitrosamine-contaminated drug and 2) generation of keratinocytic skin cancer. The pathogenesis of high-risk periocular localized basal cell carcinomas was until recently shrouded in mystery as it was mainly and until now associated with 1) intake of phototoxic drugs and 2) intense exposure to UV radiation (without intake of drugs), 3) congenital or acquired immunodeficiencies, and 4) Goltz Gorlin syndrome or 5) Xeroderma pigmentosum. Nitrosamines/ NDSRIs within the framework of polycotaminated drug intake appear to be one reasonable additional explanation for the association between carcinogen intake and subsequent skin cancer development and progression, and a relatively short-term one at that. Recently published scientific data provide information on a new ability of some of the nitrosamines - namely that some of them are photocarcinogenic or genotoxic after activation with UVA radiation. We present 4 patients who developed high-risk periocular localized basal cell carcinomas of the skin after/within the intake of potentially nitrosamine-contaminated drugs. The presented data are confirmatory with respect to previously published scientific observations on the carcinogenic effects of valsartan, candesartan, bisoprolol, metoprolol, perindopril, lisinopril and amlodipine. The contribution of newly validated data concerning potential/actual carcinogenic/genotoxic activity in the article is also due to the following newly announced nitroso preparations: torasemide, moxonidine and mirabegron. The expansion of the ˝bases of the pyramid˝ determining the stability of drug related (Photo) Nitrosogenesis/ Carcinogenesis (in terms of skin cancer generation) is growing daily. Exogenously/drug-induced Nitrosogenesis and the subsequently triggered carcinogenesis are a completely new explanatory concepts concerning the pathogenesis of skin tumors that remained unanalyzed and hidden for decades. Until now. The official lack of 1) availability, and of 2) precise concentrations regarding nitrosamines in medicinal preparations, are some of the most unexplained acts of irresponsibility to end-users and remain for the moment without a definitive answer from either regulators and manufacturers respectively. Polycontamination of polymedication in polymorbid patients remains highly problematic, at least as a cofactor in the development and progression of keratinocytic cancers, and this in the short term. Recently published data but also data from the past are suggestive that nitrosamines in tobacco are pivotal in the development of acquired mutations in p53 and RAS oncogenes in humans and rodents. The same genes are also affected by mutations in keratinocytic cancer patients. The overlapping mutation patterns of UV radiation-induced mutations in target genes such as p53 and RAS with those caused by some nitrosamines is indicative of a synergism available in terms of gene toxicity or possibly photocarcinogenicity of the latter. What leads the scientific community to believe that the nitrosamines in drugs, similar in composition and carcinogenic potency, act differently, is unclear. The link between drug intake, nitrosamine contamination, generation of some acquired mutations and subsequent cancer development becomes more than obvious and logically conditioned. The thesis of the controlled spread of cancer sounds more than logical today because: whoever controls and regulates the spread of carcinogens/mutagens/nitrosamines is also able to control the occurrence and spread of skin cancer. The Pharmaco-oncogenesis of skin cancer is determined by exogenously mediated Nitrosogenesis or the permissive availability for certain nitrosamines in drugs worldwide.
Topics: Humans; Torsemide; Prospective Studies; Retrospective Studies; Tumor Suppressor Protein p53; Carcinogenesis; Cell Transformation, Neoplastic; Skin Neoplasms; Carcinoma, Basal Cell; Nitrosamines; Acetanilides; Imidazoles; Thiazoles
PubMed: 38609117
DOI: No ID Found -
International Journal of Biological... May 2024Human heavy chain ferritin (HFn) protein cage has been explored as a nanocarrier for targeted anticancer drug delivery. Here, we introduced a matrix metalloproteinases...
Human heavy chain ferritin (HFn) protein cage has been explored as a nanocarrier for targeted anticancer drug delivery. Here, we introduced a matrix metalloproteinases (MMPs)-cleavable sequence into the DE loop of HFn, creating an MMP-responsive variant, MR-HFn, for localized and extracellular drug release. The crystal structure of MR-HFn revealed that the addition of the MMPs recognition sequence did not affect the self-assembly of HFn but presented a surface-exposed loop susceptible to MMPs cleavage. Biochemical analysis indicated that this engineered protein cage is responsive to MMPs, enabling the targeted release of encapsulated drugs. To evaluate the therapeutic potential of this engineered protein cage, monosubstituted β-carboxy phthalocyanine zinc (CPZ), a type of photosensitizer, was loaded inside this protein cage. The prepared CPZ@MR-HFn showed higher uptake and stronger phototoxicity in MMPs overexpressed tumor cells, as well as enhanced penetration into multicellular tumor spheroids compared with its counterpart CPZ@HFn in vitro. In vivo, CPZ@MR-HFn displayed a higher tumor inhibitory rate than CPZ@HFn under illumination. These results indicated that MR-HFn is a promising nanocarrier for anticancer drug delivery and the MMP-responsive strategy here can also be adapted for other stimuli.
Topics: Humans; Antineoplastic Agents; Protein Engineering; Drug Liberation; Matrix Metalloproteinases; Animals; Cell Line, Tumor; Mice; Ferritins; Indoles; Drug Carriers; Photosensitizing Agents
PubMed: 38604418
DOI: 10.1016/j.ijbiomac.2024.131492 -
International Journal of Pharmaceutical... 2024Alopecia is a chronic dermatological disorder that affects patients worldwide, with a significant impact on quality of life, self-esteem, and psychological wellbeing.... (Randomized Controlled Trial)
Randomized Controlled Trial
Alopecia is a chronic dermatological disorder that affects patients worldwide, with a significant impact on quality of life, self-esteem, and psychological wellbeing. However, commercially available options for alopecia treatment are still limited. Considering that topical formulations have a long-term use therapeutic profile, the safety of their ingredients should be closely evaluated to avoid potentially irritant substances. Alternative active ingredients with different mechanisms of action, as well as adequate vehicles, might increase patients' adherence leading to better clinical outcomes. The purpose of this study was to examine the irritation, skin sensitization, photoallergy, and phototoxicity potential of a line of ready-to-use vehicles for producing topical therapies for alopecia treatments, TrichoConcept™. Subjects were selected and randomly assigned to compare the patch test with the study products or to the control solution (sterile 0.9% NaCl solution). No clinical signs of irritation, sensitization, photoallergy or phototoxicity were reported. From the results of this study, it is suggested that the investigated products can be considered safe under the evaluated conditions, and the claims "dermatologically tested", "clinically tested", and "nonirritant" can be supported.
Topics: Humans; Dermatitis, Photoallergic; Precision Medicine; Quality of Life; Skin; Alopecia; Cosmetics
PubMed: 38604145
DOI: No ID Found -
Biomaterials Jul 2024Fungi infection is a serious threat to public health, but an effective antifungal strategy remains a challenge. Herein, a biomimetic nanocomposite with...
Fungi infection is a serious threat to public health, but an effective antifungal strategy remains a challenge. Herein, a biomimetic nanocomposite with multifunctionalities, including fungi diagnosis, antifungal adhesion, precise fungi elimination, and cytokine sequestration, is constructed for battling Candida albicans (C. albicans) infection. By screening a range of cells, we find that the polarized macrophage cells have the strongest binding tendency toward C. albicans. Thus, their membranes were exfoliated to camouflage UCNPs and then decorated with photosensitizers (methylene blue, MB) and DNA sensing elements. The resulting nanocomposite can tightly bind to fungal surfaces, promote DNA recognition, and squeeze pro-inflammatory cytokines to relieve inflammation. Consequently, this nanocomposite can detect C. albicans with enhanced sensitivity and precisely eliminate fungal cells through photodynamic therapy with minimal phototoxicity because of its switchable fluorescence behavior. The developed nanocomposite with good biocompatibility achieves a satisfactory diagnostic and therapeutic effect in a C. albicans-infected mouse model, which offers a unique approach to fight fungi infection.
Topics: Animals; Candida albicans; Nanocomposites; Mice; Biomimetic Materials; Candidiasis; Theranostic Nanomedicine; Antifungal Agents; RAW 264.7 Cells; Photochemotherapy; Photosensitizing Agents; Mice, Inbred BALB C; Biomimetics; Humans; Methylene Blue
PubMed: 38603827
DOI: 10.1016/j.biomaterials.2024.122561 -
Discover Nano Apr 2024Modern imaging strategies are paramount to studying living systems such as cells, bacteria, and fungi and their response to pathogens, toxicants, and nanomaterials (NMs)... (Review)
Review
Modern imaging strategies are paramount to studying living systems such as cells, bacteria, and fungi and their response to pathogens, toxicants, and nanomaterials (NMs) as modulated by exposure and environmental factors. The need to understand the processes and mechanisms of damage, healing, and cell survivability of living systems continues to motivate the development of alternative imaging strategies. Of particular interest is the use of label-free techniques (microscopy procedures that do not require sample staining) that minimize interference of biological processes by foreign marking substances and reduce intense light exposure and potential photo-toxicity effects. This review focuses on the synergic capabilities of atomic force microscopy (AFM) as a well-developed and robust imaging strategy with demonstrated applications to unravel intimate details in biomedical applications, with the label-free, fast, and enduring Holotomographic Microscopy (HTM) strategy. HTM is a technique that combines holography and tomography using a low intensity continuous illumination laser to investigate (quantitatively and non-invasively) cells, microorganisms, and thin tissue by generating three-dimensional (3D) images and monitoring in real-time inner morphological changes. We first review the operating principles that form the basis for the complementary details provided by these techniques regarding the surface and internal information provided by HTM and AFM, which are essential and complimentary for the development of several biomedical areas studying the interaction mechanisms of NMs with living organisms. First, AFM can provide superb resolution on surface morphology and biomechanical characterization. Second, the quantitative phase capabilities of HTM enable superb modeling and quantification of the volume, surface area, protein content, and mass density of the main components of cells and microorganisms, including the morphology of cells in microbiological systems. These capabilities result from directly quantifying refractive index changes without requiring fluorescent markers or chemicals. As such, HTM is ideal for long-term monitoring of living organisms in conditions close to their natural settings. We present a case-based review of the principal uses of both techniques and their essential contributions to nanomedicine and nanotoxicology (study of the harmful effects of NMs in living organisms), emphasizing cancer and infectious disease control. The synergic impact of the sequential use of these complementary strategies provides a clear drive for adopting these techniques as interdependent fundamental tools.
PubMed: 38594446
DOI: 10.1186/s11671-024-04003-x -
Cureus Mar 2024Introduction Skin photoaging is caused by prolonged exposure to sunlight, particularly ultraviolet rays (UV). High cumulative levels of UV radiation may cause burning,...
Introduction Skin photoaging is caused by prolonged exposure to sunlight, particularly ultraviolet rays (UV). High cumulative levels of UV radiation may cause burning, photoallergic or phototoxic reactions, pigmentary changes, photoaging, and even immunosuppression and skin cancers. Therefore, this study aims to assess knowledge, attitude, reception, and preventive practices towards skin photoaging among the Jazan general population in Saudi Arabia and its determinants. Methods A descriptive cross-sectional study was conducted among the general population of Jazan, Saudi Arabia, who were aged 18 years and above and agreed to participate in the study. The calculated minimum sample size was 385. An online, semi-structured, self-administered questionnaire was distributed conveniently in Google Forms through social media platforms. It included four sections: The first section was about sociodemographic characteristics. The second section assessed the smoking, exercise, and healthy diet behavior of participants and the use of sunscreen. The third section assessed the knowledge regarding the photoaging process and its preventive measures utilization using three-point Likert scale questions. The fourth section assessed attitudes towards the photoaging process and its preventive measures through three-point Likert scales. Results The study included 452, of which 243 (53.76%) were aged 18-30 years, 258 (57.08%) were females, and 272 (60.18%) had white skin color. Approximately 417 (92.26%) were nonsmokers. Sixty-eight percent (372) spent 1-3 hours in the sun. Social media was the primary source of information on photoaging 81 (17.92%). Around 234 (51.77%) defined photoaging correctly. Regarding sunscreen usage, 58 (12.83%) always use sunscreen, and 177 (39.16%) never use it. However, 191 (42.26%) recognized the correct sunscreen application. Approximately 233 (51.5%) and 240 (53.1%) of respondents had fair knowledge and a positive attitude regarding photoaging and sunscreen use. Being female, pursuing university and postgraduate education, and taking information on photoaging from a physician were linked to a higher knowledge of photoaging (p<0.05). Participants who never use sunscreen had lower knowledge than those who always use it (p<0.001). None of the demographic factors was associated with the attitude towards sunscreen use (p>0.05). Conclusion There is a substantial gap in knowledge and preventive practices related to skin photoaging among the Jazan general population in Saudi Arabia. Gender, education level, and information sources influence knowledge levels. Targeted educational interventions are needed to enhance awareness and promote healthier practices, particularly sun exposure and photoaging prevention.
PubMed: 38586780
DOI: 10.7759/cureus.55710 -
Journal of Inorganic Biochemistry Jul 2024Trinuclear ruthenium(II) polypyridyl complexes anchored to benzimidazole-triazine / trisamine scaffolds were investigated as photosensitizers for photodynamic therapy....
Trinuclear ruthenium(II) polypyridyl complexes anchored to benzimidazole-triazine / trisamine scaffolds were investigated as photosensitizers for photodynamic therapy. The trinuclear complexes were noted to produce a significant amount of singlet oxygen in both DMF and aqueous media, are photostable and show appreciable emission quantum yields (ɸ). In our experimental setting, despite the moderate phototoxic activity in the HeLa cervical cancer cell line, the phototoxic indices (PI) of the trinuclear complexes are superior relative to the PIs of a clinically approved photosensitizer, Photofrin®, and the pro-drug 5-aminolevulinic acid (PI: >7 relative to PI: >1 and PI: 4.4 for 5-aminolevulinic acid and Photofrin®, respectively). Furthermore, the ruthenium complexes were noted to show appreciable long-term cytotoxicity upon light irradiation in HeLa cells in a concentration-dependent manner. Consequently, this long-term activity of the ruthenium(II) polypyridyl complexes embodies their ability to reduce the probability of the recurrence of cervical cancer. Taken together, this presents a strong motivation for the development of polymetallic complexes as anticancer agents.
Topics: Humans; Photosensitizing Agents; HeLa Cells; Uterine Cervical Neoplasms; Ruthenium; Female; Coordination Complexes; Photochemotherapy; Antineoplastic Agents; Pyridines; Singlet Oxygen
PubMed: 38581803
DOI: 10.1016/j.jinorgbio.2024.112545